Sesquiterpenes from Chloranthus holostegius with anti-inflammatory activities.

The phytoconstituents of the whole plants of Chloranthus holostegius were investigated. As a result, thirteen undescribed sesquiterpenes (chloranholosins A-M, 1-13), including ten acorane-type sesquiterpenes (1-10), one germacrene-type sesquiterpene (11), and two lindenane-type sesquiterpenes (12-13), together with fifteen known sesquiterpenes were isolated. Their structures and absolute configurations were elucidated by a comprehensive method including the spectroscopic data, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction. Chloranholosin L (12) was elucidated as a rare lindenane-type sesquiterpene featuring 14α-Me and 5-OH moieties. And chloranholosin M (13) was the first lindenane-type sesquiterpene possessing ß-cyclopropane, 14α-Me, and 5ß-H configuration from the family Chloranthaceae. Furthermore, twelve new isolates and some known sesquiterpenes were evaluated for their inhibitory activity against LPS-induced nitric oxide (NO) production in RAW 264.7 macrophage cells. Among them, compounds 12, 16, and 23 showed comparable inhibitory activity to that of the positive control, with IC50 values of 47.9, 41.5, and 48.3 µM, respectively.

Photo induced reaction of myoglobins with energy transferred from excited free tryptophan.

Despite being one of the most studied proteins in biology, the photolysis mechanism of myoglobin heme affected by endogenous substances free amino acids is still in controversy. The transient absorption and kinetic processes of photo-excited myoglobin in three forms and the effects of free excited tryptophan on redox reaction of myoglobin were monitored by laser flash photolysis. With dual energy superposition of direct light irradiation and indirect energy transferred from the free excited tryptophan, the variation value in optical density (ΔOD) of MetMb increased by 66.7%, from 0.9 to 1.5. The ΔOD value of MbO2 in ferrous form increased from 0.9 to 1.25, while the ΔOD value of DeoxyMb increased from 0.75 to 1.2. The decay time of excited DeoxyMb was prolonged obviously with the excited tryptophan, while the decay time of excited MbO2 and MetMb was shortened significantly. The excited tryptophan could promote laser induced reaction processes of myoglobin in different forms by intermolecular energy transfer to one final similar photo reaction state. The possible photo induced reaction mechanisms of DeoxyMb, MbO2, MetMb with and without free tryptophan were also proposed.

Draft Genome Sequence of Bacillus farraginis R-6540T (DSM 16013), a Spore-Forming Bacterium Isolated at Dairy Farms.

Bacillus farraginis R-6540(T) is a Gram-positive, aerobic, and spore-forming bacterium with very high intrinsic heat resistance. Here, we report the 5.32-Mb draft genome sequence of B. farraginis R-6540(T), which is the first genome sequence of this species and will promote its fundamental research.

Draft Genome Sequence of Bacillus shackletonii LMG 18435T, Isolated from Volcanic Mossy Soil.

Bacillus shackletonii LMG 18435(T) is a Gram-positive, aerobic, and spore-forming bacterium. Here, we report the 5.30-Mb draft genome sequence of B. shackletonii LMG 18435(T), which will promote its fundamental research and provide useful information for genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Draft Genome Sequence of Bacillus plakortidis P203T (DSM 19153), an Alkali- and Salt-Tolerant Marine Bacterium.

Bacillus plakortidis P203(T) is a Gram-positive, spore-forming, and alkali- and salt-tolerant marine bacterium. Here, we report the 3.97-Mb draft genome sequence of B. plakortidis P203(T), which will promote its fundamental research and provide useful information for genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Draft Genome Sequence of Bacillus muralis LMG 20238T (DSM 16288), a Spore-Forming Bacterium Isolated from Deteriorated Mural Paintings.

Bacillus muralis LMG 20238(T) is a Gram-positive, aerobic, and spore-forming bacterium. Here, we report the 5.18-Mb draft genome sequence of B. muralis LMG 20238(T), which is the first genome sequence of this species and will promote its fundamental research.

Surface Molecularly Imprinted Polymer of Chitosan Grafted Poly(methyl methacrylate) for 5-Fluorouracil and Controlled Release.

The molecular surface imprinted graft copolymer of chitosan with methyl methacrylate (MIP-CS-g-PMMA) were prepared by free radical polymerization with 5-fluorouracil (5-FU) as the template molecule using initiator of ammonium persulfate as adsorption system. MIPs were characterized by FTIR, X-ray diffraction, thermo-gravimetric analysis, (1)H NMR and SEM. The mechanism of graft copolymerization and factors affected graft reaction were studied in details, and the optimum reaction conditions (to the highest %G and %E as the standard) were obtained at [MMA] 1.2 mol/L, [Chitosan] 16.67 mol/L, [initiator] 0.0062 mol/L, temperature 60 °C and reaction time 7 h. MIPs exhibited high recognition selectivity and excellent combining affinity to template molecular. The in vitro release of the 5-FU was highly pH-dependent and time delayed. The release behavior showed that the drugs did not release in simulated gastric fluid (pH = 1.0), and the drug release was small in the simulated small intestinal fluid (pH = 6.8), and drug abrupt release will be produced in the simulated colon fluid (pH = 7.4), indicating excellent colon-specific drug delivery behavior.

High-Quality Draft Genome Sequence of Aneurinibacillus migulanus ATCC 9999T (DSM 2895), a Gramicidin S-Producing Bacterium Isolated from Garden Soil.

Aneurinibacillus migulanus ATCC 9999(T) (DSM 2895) is a Gram-positive, round-spore-forming, and gramicidin S-producing bacterium. Here, we report the 6.35-Mb high-quality draft genome sequence of A. migulanus ATCC 9999(T), which will provide useful information for the genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Genome Sequence of Bacillus butanolivorans K9T (DSM 18926), an n-Butanol-Consuming Bacterium Isolated from Soil.

Bacillus butanolivorans K9(T) (DSM 18926) is a Gram-positive, spore-forming, strictly aerobic, and n-butanol-consuming bacterium. Here, we report the 5.68-Mb genome sequence of B. butanolivorans K9(T), which is the first genomic information of this species that will provide useful information for the genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Draft Genome Sequence of Bacillus murimartini LMG 21005T, an Alkalitolerant Bacterium Isolated from a Church Wall Mural in Germany.

Bacillus murimartini LMG 21005(T) is a Gram-positive, spore-forming, and alkalitolerant bacterium isolated from a church wall mural. Here, we report the 4.17-Mb genome sequence of B. murimartini LMG 21005(T), which will accelerate the application of this alkalitolerant bacterium and provide useful information for genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Draft Genome Sequence of Sporosarcina globispora W 25T (DSM 4), a Psychrophilic Bacterium Isolated from Soil and River Water.

Sporosarcina globispora W 25(T) (DSM 4) is a Gram-positive, round-spore-forming, and psychrophilic bacterium. Here, we report the 5.66-Mb genome sequence of S. globispora W 25(T), which will accelerate the application of this psychrophile and provide useful information for genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Genome Sequence of Type Strain Bacillus decisifrondis E5HC-32T (DSM 11725T), Isolated from Soil Underlying the Decaying Leaf Litter of a Slash Pine Forest.

Bacillus decisifrondis E5HC-32(T) (DSM 11725(T)) is a Gram-positive, subterminal spherical spore-forming, strictly aerobic bacterium. Here, we report the 5,613,728-bp genome sequence of B. decisifrondis E5HC-32(T), which is the first genome information of this species and will provide useful information for genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Draft Genome Sequence of Type Strain Lysinibacillus xylanilyticus DSM 23493T.

Lysinibacillus xylanilyticus DSM 23493(T) is a Gram-positive, spore-forming bacterium. Here, we report the 5.22-Mb genome sequence of Lysinibacillus xylanilyticus DSM 23493(T), which will accelerate the application of degrading xylan and provide useful information for genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Draft Genome Sequence of Bacillus pseudalcaliphilus PN-137T (DSM 8725), an Alkaliphilic Halotolerant Bacterium Isolated from Garden Soils.

Bacillus pseudalcaliphilus PN-137(T) (DSM 8725) is a Gram-positive, spore-forming, alkaliphilic, and halotolerant bacterium. Here, we report the 4.49-Mb genome sequence of B. pseudalcaliphilus PN-137(T), which will accelerate the application of this alkaliphile and provide useful information for genomic taxonomy and phylogenomics of Bacillus-like bacteria.

Spectroscopic insights into the Photoreduction of Cytochrome c with UVA-Vis Light Irradiation.

With the particular conjugation structure in the heme prosthetic group, Cyt c shows unusual functions similar to chlorophyll while irradiated by specific wavelength of UV-Vis lights. To further reveal mechanism of the photo-irradiation of Cyt c, we then studied various external factors that may influence the photo induced process. The absorbance intensity increase of band (317 nm) and Q band (520 nm and 549 nm)indicated Cyt c in phosphate-buffered saline within N2 atmosphere was photoreduced to Fe(II) Cyt c. Irradiated by 410 nm, the photoreduction process was facilitated by Met. But Trp, Tyr and Phe impeded the process due to their light absorbance abilities. In addition, the results of fluorescence and CD spectra indicated that the microenvironment polarity of Trp residue varied during the photoreduction process. And the secondary structure of Cyt c changed with lower α-helix/ßsheet ratio. The photoreduction mechanism of Cyt c was intramolecular electron transfer and porphyrin cation radicals were generated. The protein structure of Cyt c changed as well as part of the photoreduction.

Bacillus taiwanensis sp. nov., isolated from a soil sample from Taiwan.

A Gram-stain-positive, rod-shaped, endospore-forming, aerobic bacterium (FJAT-14571(T)) was isolated from a soil sample in Taiwan. Strain FJAT-14571(T) grew at 20-40 °C (optimum 35 °C), pH 6-10 (optimum pH 8) and 0-2% (w/v) NaCl (optimum 0%). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain FJAT-14571(T) was a member of the genus Bacillus and was most closely related to Bacillus oceanisediminis DSM 24771(T) (96.2%). DNA-DNA relatedness between strain FJAT-14571(T) and B. oceanisediminis DSM 24771(T) was low (32.0% ± 0.88%). The diagnostic diamino acid of the peptidoglycan of strain FJAT-14571(T) was meso-diaminopimelic acid and the predominant menaquinone was MK-7 (96.6%). The major cellular fatty acids were iso-C15 : 0 (46.4%), anteiso-C15 : 0 (7.6%), iso-C17 : 0 (8.2%) and iso-C16 : 0 (10.0 %) and the DNA G+C content was 40.8 mol%. Phenotypic, chemotaxonomic and genotypic properties clearly indicated that strain FJAT-14571(T) represents a novel species within the genus Bacillus, for which the name Bacillus taiwanensis sp. nov. is proposed. The type strain is FJAT-14571(T) ( = DSM 27845(T) = CGMCC1.1 2698(T)).

[Spectral Study on Coordination Reaction Between metMyoglobin and Nitric Oxide].

As we all known, the instantaneous reaction between protein and ligands are very important to adjust the normal playing of biological function. And nitric oxide interactions with iron are the most important biological reactions in which NO participates. Unlike carbon monoxide or oxygen, NO can also bind reversibly to ferric iron. In this paper, UV-Vis absorption and CD spectra were used to study coordination reaction process between horse heart metMb and NO, to demonstrate the coordination reaction mechanism and to explore the influencing factors of metMb with NO. The experimental results showed that metMb could react with NO, and obtained three new peaks at 420 nm, 534 and 568 nm, respectively, which implied metMb and NO have reacted and generated a new complex-nitrosylmetmyoglobin (metMbNO). Then as time went on, NO concentration decreased in the solution, and the Fe-N bond fractured under the attack of H2O, then NO leaves slowly from metMbNO, and met-Mb was regenerated. In this experiment, we also found that external conditions such as buffer medium, ionic strength, pH, temperature, etc, had an important influence on the coordination reaction between metMb and NO. It was favorable for the coordination reaction, when the 0.01 mol x L(-1) phosphate buffer. solution is near neutral condition, the temperature is 280 K, the coordination reaction could reach equilibrium at a fastest speed. In addition, the CD date show that NO only reacts with Fe atom in the center of heme and has less effect on the secondary structuers of protein. The research of metMb and NO played an important role to further study the function of NO. Especially the establish of equilibrium reaction mechanism between NO and heme protein has an important research value on maintaining the balance of NO in vivo and keeping the normal function in the body's cells.

[DFT/TDDFT study on the structure and absorption spectra of free base, N-/neo-confused porphyrin].

Porphyrin is an important class of photochemical materials, which has been widely used in various fields. Computational investigations into the ground state structures and orbital energy levels of free base porphyrin (FBP), neo-confused porphyrin (NECP) and N-confused porphyrin (NCP) were performed with density functional theory(DFT). Absorption spectra were calculated at TD-B3LYP/6-31+G(d). Degeneracy of HOMO and HOMO-1 is lost, which would account for the shoulder peaks about the most intense transitions of FBP and NECP. Following FBP, NECP and NCP order, the orbital energy level (OEL) of LUMO decreases while the OEL of HOMO increases, which lead to the red shift of adsorption spectra. The energy difference between LUMO and LUMO+1 is almost the same as the difference between HOMO and HOMO-1, which would account for the only most intense transition of NCP. Solvent effect on ground state structures and absorption spectra was also investigated. The data shows that the character peak of Soret band and Q band changes in different solvent (benzene, chloroform, acetonitrile and water). So we further focus on discussing the N atom position and solvent effects on the energy level and Soret/Q bands of FBP, NCP and NECP, as well as clarifying its variation regularity and mechanism.

[Spectroscopic studies of guanidine hydrochloride-induced unfolding of hemoglobin].

In the present paper, based on the ultraviolet-visible (UV-Vis) absorption spectroscopy, fluorescence spectroscopy, and stopped flow-fluorescence spectroscopy, the authors studied the protein unfolding process of hemoglobin induced by GdmHcl. The experiments result shows that there were two different procedures about GdmHcl inducing hemoglobin unfolding from the evidences of UV-Vis absorption spectrum and fluorescence phase diagrams. Namely, the hemoglobin subunit exhibits depolymerization, forming the intermediates when incubated with GdmHcl at the concentration of 1. 0 mol x L(-1). With the increase in the concentration, various subunit structure became loose gradually, and the protoheme collapsed eventually. UV-Vis absorption spectroscopy indicates that the addition of reductant can cooperate with the depolymerization of hemoglobin subunit and the disaggregation of protoheme. The reductant results in the unfolding procedure that hemoglobin from "three-state model" turns into "two-state model".

[Comparison between myoglobin and its mutant(D60K) interacting with hydrogen peroxide by spectrum].

To characterize the roles played by surface-charged residue Asp60 in the structure stability of myoglobin when it was replaced with Lys, the interaction of myoglobin[Mb(WT)] and its mutant[Mb(D60K)] with hydrogen peroxide (H2O2) were studied by the method of ultraviolet-visible (UV-Vis) absorption spectroscopy, fluorescence spectroscopy and stopped-flow fluorescence spectroscopy under simulative physiological conditions. There are remarkable differences between Mb(D60K) and Mb (WT) in the UV-Vis absorption spectroscopy and fluorescence spectroscopy of iron porphyrin during the process of interaction. Although we only altered one external amino acide, the data showed that the function and structure stability of Mb(D60K) was greatly changed. Furthermore, results from synchronous fluorescence spectroscopy and stopped-flow fluorescence spectroscopy all indicated that H2O2 had less effect on the structure of Mb(D60K) while the structure of Mb(WT) was notably changed. From a comprehensive and comparative data analysis, the authors determined that the structure of Mb(D60K) was improved when it interacted with H2O2.